The subject matter of the present invention relates to nucleoside derivatives with photolabile protective groups, a method for the production thereof, their use and nucleic acid chips built up therefrom.
Photolabile protective groups are of significance for the hydroxy and phosphate functions in nucleosides and/or nucleotides since they are suitable for the light-controlled parallel synthesis of oligonucleotides on a solid support surface (Fodor et al., Science 1991, 251, p. 767 et seq.). It is possible to synthesize therewith oligonucleotides or nucleic acid chips which can be used for sequencing nucleic acids efficiently, for example.
To date, it is only known to produce DNA chips by means of photolithographic methods using 3xe2x80x2-O phosphite amides, which correspondingly carry the temporary photolabile protective group at the 5xe2x80x2-O position (WO-A-96/18634). It is possible to produce DNA chips by means of these nucleic acid building blocks, the oligomer being built up from the 3xe2x80x2 end to the 5xe2x80x2 end. The finished oligomer is thus anchored to the solid phase via the 3xe2x80x2-O end. The 5xe2x80x2-OH end can be accessed freely. DNA chips which were produced with this method can be used for hybridization experiments but not for certain enzyme reactions (e.g. with DNA polymerase or ligase) which require free 3xe2x80x2-OH.
It is the object of this invention to provide 3xe2x80x2-photolabile nucleosides and their derivatives and to generate nucleic acid chips using the 3xe2x80x2-photolabile nucleosides obtained therefrom, in which the oligomers built up by light-controlled synthesis are linked to the solid phase via the 5xe2x80x2 end and thus enable enzyme reactions at the 3xe2x80x2 end.